Bioluminescence tomography (BLT) we invented in 2002 is a rapidly developing optical molecular imaging modality for small animal imaging. The state-of-the art results have indicated that BLT can indeed produce valuable tomographic information in favorable cases or with strong prior knowledge. However, the primary challenge in this area remains that it is highly desirable but extremely difficult to stabilize and improve the current BLT performance. It has been recently reported that bioluminescent spectra are significantly affected by temperature, which we recognize as a major opportunity to transform the ill-posed BLT problem into a well-posed setting. Specifically, with focused ultrasound heating, a bioluminescent source distribution in a mouse can be precisely perturbed in a local and controllable fashion. The resultant difference in the externally captured optical signals is solely related to the source parameters in the heated region. Our hypothesis is that this temperature dependence of the bioluminescence can be utilized in the BLT reconstruction for superior image quality. Our overall goal is to develop temperature-modulated bioluminescence tomography (TBT) for mouse studies. The three specific aims are to (1) prototype a focused ultrasound array to heat one small volume of interest (VOI) each time in a living mouse, (2) develop a BLT algorithm to reconstruct an underlying source distribution based on the bioluminescent data measured before and after heating, (3) evaluate the TBT technology in numerical simulation and phantom experiments, as well as apply and validate it in mouse studies. Upon the completion of this project, we will have built the first TBT system and demonstrated its in vivo validity and utility. This success will represent a major step forward in the field of BLT. Since bioluminescent imaging has been used in almost all the mouse models of human diseases, our proposed TBT technology shall have a general, important and immediate applicability in mouse studies, and for development of molecular medicine. Bioluminescence tomography (BLT) invented by us in 2002 is to localize and quantify bioluminescent sources in studies of small animals as many human disease models. This project will improve significantly the accuracy and robustness of the BLT techniques by means of focused ultrasound heating to induce bioluminescent signal changes on the mouse body surface. Our technology shall have a general, important and immediate applicability in mouse studies, and facilitate the development of molecular medicine. [unreadable] [unreadable] [unreadable]